Viral capsid proteins and any peptides or compositions thereof for the treatment of pathologic disorders

ABSTRACT

The present invention relates to viral capsid proteins, as a medicament for the treatment of a pathologic disorder. More particularly, the invention relates to the viral capsid proteins VP1, VP2 and VP3, preferably, the SV40 VP1 or any peptide, fragment, mutant, derivative and mixtures thereof or of virus-like particles (VLP&#39;s) comprising the same, as the active ingredient in compositions for the treatment of pathologic disorders, preferably disorders associated with inactivation of cellular proteins involved with quality control processes, particularly, chaperones. The invention further provides methods for the treatment of such disorders and the use of the SV40 capsid proteins for the preparation of pharmaceutical compositions.

FIELD OF THE INVENTION

The present invention relates to compositions and methods for treatingdisorders associated with inactivation of cellular proteins involvedwith quality control processes. More particularly, the invention relatesto the use of viral capsid proteins, preferably, the SV40 VP1 or anypeptide, fragment, mutant and mixture thereof in compositions andmethods for the treatment of such pathologic disorders.

BACKGROUND OF THE INVENTION

All publications mentioned throughout this application are fullyincorporated herein by reference, including all references citedtherein.

Viruses, the ultimate parasites, usurp many cellular functions for theirmultiplication [Munther et al., Science's STKE 335:1-13 (2006)]. As soonas they bind to cells they trigger multiple cascades of events that theyemploy for cell entry, trafficking and disassembly, before they startutilizing cellular machinery for expression and replication. Some of thefactors activated by the infecting virus participate in key processessuch as inflammatory response and cell death.

The very early events that occur before disassembly and expression ofthe viral genome are most likely triggered by the viral outer shell.Thus viral capsids may be used to activate cellular mechanisms that mayhave therapeutic effects.

SV40 (and other members of the polyomavirus and papilloma virusfamilies) induce chaperones following infection [Ioannis et al., FEBSLetters 355:282-286 (1994); Cripe et al., J. Virol. 69:7807-7813 (1995);Chromy et al., PNAS 100:10477-10482 (2006) and references therein],presumably because they utilize host chaperons for disassembly (andassembly). As surprisingly shown by the present invention, chaperonesare induced by the capsid proteins, or the viral structural proteins,rather than by viral regulatory proteins. Chaperons were proposed toameliorate critical conditions such as ARDS (acute respiratory distresssyndrome) and AKI (Acute Kidney Injury), which was previously referredto as ATN (acute tubular necrosis). In particular, ectopic expression ofHSP70, applied by gene therapy to model ARDS rats, showed amelioratingeffect [Weiss et al., the J. Clin. Invest. 110:801-806 (2002)].Therefore, the inventors examined the possibility that induction ofchaperones by SV40 may serve to treat these conditions. As clearly shownby the present invention, SV40 capsids (VLPs) remarkably amelioratedpathological symptoms using the AKI mice model, and are thereforeapplicable for use in the treatment of Acute Renal Failure (ARF). Thisparticular example (ARF) clearly establishes the feasibility of usingviral capsid proteins, and specifically the SV40 VP1, for treating anyother disorders associated with disfunction of cellular quality controlmechanisms, in variety of organs.

It is therefore one object of the invention to provide viral capsidproteins VP1, VP2 and VP3, preferably, the SV40 VP1 or any peptide,mutant, fragment and mixture thereof as the active ingredient incompositions for use in the treatment of pathologic disorders,preferably disorders associated with inactivation of cellular proteins.Such cellular proteins are preferably proteins involved with qualitycontrol processes, for example, chaperones.

In yet another object the invention provides methods for the treatmentof disorders associated with inactivation of cellular proteins,preferably proteins involved with quality control processes, forexample, chaperones.

Another object of the invention is to provide methods for enhancing theameliorating effect of cellular proteins involved with quality controlprocesses, for example, chaperones, on pathologic disorders, byaugmenting these cellular proteins.

In another object, the invention provides the use of SV40 capsidproteins, or SV40 VLP's comprising said capsid proteins, in thepreparation of compositions for the treatment of pathologic disorders,preferably, immune-related disorders or neurodegenerative disorders.

These and other objects of the invention will become apparent as thedescription proceeds.

SUMMARY OF THE INVENTION

In a first aspect the present invention relates to a pharmaceuticalcomposition for the treatment of a pathologic disorder. The compositionof the invention comprises as an active ingredient a therapeuticallyeffective amount of at least one viral capsid protein or any peptide,mutant or fragment thereof, or any VLP's comprising at least one capsidprotein or any peptide, mutant or fragment thereof.

It should be noted that the composition of the invention, althoughintended for the treatment of pathologic disorders, is not intended tobe used as a vaccine against the virus in which said capsid proteinsderived from.

According to one embodiment, the viral capsid protein may be any capsidproteins of papillomaviruses or polyomaviruses, or any fragment,peptide, mutant, any mixture and combinations thereof.

According to a specifically preferred embodiment, the viral capsidprotein may be at least one of SV40 VP1, VP2, VP3, and any peptide,mutant, fragment, any mixture and any combinations thereof or any VLP'scomprising at least one of said SV40 capsid proteins or any peptide,mutant or fragment thereof. Most preferably, the viral capsid proteinmay be SV40 VP1 or any peptide, mutant or fragment thereof. Particularexample for a mutated SV40VP1 molecule may by the VP1ΔC mutant,preferably, said mutated molecule comprises the amino acid sequencesubstantially as denoted by SEQ ID NO. 4.

According to another embodiment, the composition of the invention may beparticularly applicable and suitable for the treatment of a pathologicdisorder such as a neurodegenerative disorder or an immune-relateddisorder. It should be noted that the pathologic disorder to be treatedby the composition of the invention is preferably a disorder associatedwith inactivation of cellular proteins involved with quality controlprocesses within the cell, preferably, a disorder associated withinactivation of chaperones.

In a second aspect, the present invention relates to a method for thetreatment of a pathologic disorder in a subject in need thereof. Thismethod comprises the step of administering to the treated subject atherapeutically effective amount of at least one viral capsid protein orany peptide, mutant or fragment thereof or any VLP's comprising thesame, or of a composition comprising the same.

In a third aspect, the invention relates to a method for enhancing theameliorating effect of cellular proteins participating on qualitycontrol processes, on a pathologic disorder. This method comprises thesteps of contacting cells obtained from a subject suffering of apathologic disorder with an effective amount of at least one viralcapsid protein or any peptide, mutant or fragment thereof, or of VLP'scomprising the same.

In a fourth aspect, the invention relates to the use of at least oneviral capsid protein or any peptide, mutant or fragment thereof or VLP'scomprising the same, in the preparation of a pharmaceutical compositionfor the treatment of a pathologic disorder.

The invention will be further described on hand of the following figuresand examples.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A-1C. Production of recombinant VLPs in insect cells

(A) VLPs and (B) wild-type SV40, viewed under the electron microscopefollowing staining in 1% phospho tungstate, pH 7. (C) PAGE analysis ofthe purified VLPs, stained by Coomassie. Abbreviations: Prot. (protein).

FIG. 2A-2B. SV40 VLPs up regulate Hsp/c70 in cultured CV1 cells

CV1 cells were treated with SV40 VLPs, 50 ng and 500 ng per 10⁶ cells.(A) Total cell proteins were harvested at different time points, shownin hours above, and analyzed by Western blotting with monocloncalantibodies against hsp/hsc70 proteins. Controls: HS—CV1 cells, heatshocked for 90 min at 45° C., C—untreated control. (B) CV1 cells treatedwith 50 ng VLPs, immunostained with anti-Hsp/c 70 antibody and viewedunder confocal microscopy. Abbreviations: moc. (mock).

FIG. 3A-3C. Embryonic human kidney (HEK) cells

Cells were treated with etoposide at the time of infection with VLPs, ata final concentration of 20 μg/ml. The concentration of VLPs was 50 ngper 10⁶ cells. (A) shows control; (B) shows treatment with etoposide and(C) shows treatment with etoposide and SV40 VPL's. Abbreviations: cont.(control).

FIG. 4A-4B. SV40 targets the kidney in mice

A section through the kidney of a mouse 48 hours following injectionwith 10⁸ infectious units (pfu) of SVluc vector [Arad, U. Virology304:155-159 (2002)] in 100 μl volume via the tail vein.Immunohistochemistry was performed on renal tissue with an anti-SV40 VP1[Sandalon and Oppenheim, Virology 237:414-421 (1997)] as the primaryantibody and an HRP-labeled anti-rabbit secondary antibody. Brown stainindicates VP1 uptake by renal tubular epithelial cells. PBS injectedcontrols did not show staining. (A) shows 10× magnification and (B)shows 40× magnification.

FIG. 5. Mouse model of nephrotoxicity

HgCl₂, at the designated concentration, was administered IP to BALB/Cfemale mice, blood was drawn from the tail-vein on the designated dayand urea levels were measured using Reflotron urea test (Roche). Thenumber of animals in each group is shown in parentheses. The barsrepresent standard deviation. Abbreviations: bl. Ur. (blood urea), sh.(sham).

FIG. 6. Protection of cultured mouse tubular kidney cells against HgCl₂insult

Cultured mouse tubular kidney cells were treated with HgCl₂ (A) or HgCl₂and SV40 VLP's (B), and viewed under the microscope.

FIG. 7A-7C. Survival rate of mice treated with SV40 VLP's

(A) The figure shows that treatment with VLPs increases the survivalrate of AKI model mice (No vec.=HgCl₂ only; VLP=VLP+HgCl₂); (B) showssurvival of the VLP-treated vs, untreated AKI animals; (C) shows doseresponse. Note log scale of the VLP dose. The numbers below the pointsdesignate the number of animals at the particular dose. Abbreviations:sur. (survival), D (days post HgCl₂ injection), sh. (sham).

FIG. 8A-8B. Mutated SV40 VP1 capsid protein, VP1ΔC protects AKI mice

(A) Production of the mutant protein (a), (b)—samples from two differentbatches of VP1ΔC that were injected to mice. PAGE was run in MES bufferand proteins detected by silver staining. M-SeeBlue size marker(Invitrogen). (B) Survival experiments. Abbreviations: Prot. (protein),Sur. (survival), sh. (sham).

FIG. 9A-9C. SV40 VLP's protective effect in different disease parameters

(A) Kidney gross appearance as photographed 3 days after HgCl₂injection. (B) Serum urea (left) and creatinine (right) levels weremeasured on day 4 using Reflotron kit for urea and creatinine tests(Roche) respectively. Note that serum creatinine in the sham andVLP-only treated groups was below detection level. Results of one-tailedStudent's t-Test were P=0.0008 for blood urea and P=0.035 forcreatinine. The numbers of animals in each group is designated below.(C) Serum urea of VLP-treated AKI and non-treated AKI mice. The numberof animals in each group is shown in parentheses. Bars representstandard error. Abbreviations: N. an (number of animals), D. (days), sh.(sham).

FIG. 10. Mercury-induced oxidative stress

Levels of thiobarbituric acid reactive substances were measured usingTBARS assay [Esterbauer, H. and Cheeseman, K. H. Methods Enzymol.186:407-421 (1990)]. Mice were injected with 6.5 mg/kg HgCl₂, with orwithout treatment with 0.3 mg/kg VLPs. The number in each group is shownbelow the bars. Abbreviations: N. an (number of animals), sh. (sham).

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present invention relates to a pharmaceuticalcomposition for the treatment of a pathologic disorder. The compositionof the invention comprises as an active ingredient a therapeuticallyeffective amount of at least one viral capsid protein or any peptide,mutant or fragment thereof or of VLP's comprising at least one capsidprotein or any peptide, mutant or fragment thereof.

It should be noted that the composition of the invention, althoughintended for the treatment of pathologic disorders, is not to be used asa vaccine against the virus in which said capsid proteins derived from.Therefore, the composition of the invention is for the treatment of apathologic disorder, provided that, this composition is not used as avaccine or for vaccination of the treated subject.

According to one embodiment, the viral capsid protein comprised as anactive ingredient may be any one of the capsid proteins ofpapillomaviruses or polyomaviruses, or any fragment, peptides andmixtures and combinations thereof.

The papilloma, polyoma, and vacuolating agents seem to form a naturalgroup of tumor viruses, for which the name papova virus group was used.Historically, the name papova was derived from Rabbit PApilloma virus,Mouse POlyoma virus, Simian VAcuolating virus. Members of the group areall non-enveloped viruses having double stranded DNA genome, whichperform multiplication within the cell nucleus. Papilloma andpolyomaviruses are related virus families with a common capsidarchitecture. For both, 72 capsomeres, each a pentamer of the majorcapsid protein, form a T=7 icosahedral lattice. This structuralsimilarity exists despite a complete lack of sequence homology betweenpolyomavirus (VP1) and papillomavirus (L1) major capsid proteins.

Seventy strains of Human Papillomavirus (HPV) have been identified sofar. These viruses are known for their role in causing warts (bothcommon warts and genital warts) as well as their association withcancer. Most people are infected with some strain of HPV in their lives.Two structural proteins form the capsids of papillomaviruses. The majorstructural protein L1 is the structural determinant of the capsids andis present in 360 copies arranged in 72 pentamers. The minor structuralprotein L2 is estimated to be present in 12 copies per capsid. Possibleroles for L2 in interaction with cell surface receptors and in virionuptake have been suggested. L2 localizes in sub-nuclear domainsidentified as nuclear domain 10 (ND10).

Therefore, according to a particular embodiment, the papillomaviruscapsid proteins L1 and L2 may be used as an active ingredient for thecomposition of the invention. The Polyomaviridae family of small,nonenveloped, icosahedral DNA viruses may be best represented by themurine polyomavirus, the simian virus 40 (SV40), human BK virus andhuman JC virus. Like most DNA viruses, polyomavirus capsid proteins aresynthesized in the cytosol, whereas assembly of virions occurs only inthe nucleus. Polyomavirus capsids are comprised of 72 pentamers(capsomeres) of the major capsid viral protein (VP1), which is arrangedin a T=7 icosahedral lattice ≈50 nm in diameter. One minor capsidprotein, either VP2 or VP3, binds in the central 5-fold cavity of eachVP1 either VP2 or VP3, binds in the central 5-fold cavity of each VP1pentamer. The atomic structure of the virion reveals that the C-terminaldomain of each VP1 monomer “invades” a neighboring pentamer to form theprincipal interpentamer contacts, and these contacts are stabilized bycalcium ions.

According to a specifically preferred embodiment, the viral capsidprotein may be at least one of SV40 VP1, VP2, VP3, and any peptide,mutant, fragments, mixtures and combinations thereof or VLP's comprisingat least one of SV40 VP1, VP2, VP3, or any peptide, mutant or fragmentthereof. Most preferably, the viral capsid protein may be SV40 VP1 orany peptide, mutant or fragment thereof. It should be noted thatpreferably the term SV40 VP1 protein refers to the VP1 protein havingthe amino acid sequence as indicated by GenBank Accession numberNP_(—)043126 GI: 96 28426, which is incorporated herein by reference.According to a specifically preferred embodiment, the SV40 VP1 capsidprotein comprises the amino acid sequence as denoted by SEQ ID NO:1.

SV40 is a small, double-stranded DNA Primate polyomavirus with a minichromosome of 5.2 kb. The viral capsid is composed of threeviral-encoded proteins, VP1, VP2, and VP3, that encloses theminichromosome. Encapsidation occurs by gradual addition andorganization of capsid proteins around the chromatin. The viral proteinsVP2 and VP3 are thought to bridge between the viral chromatin and theVP1 capsid shell. A domain near the carboxyl-terminus of VP3 has beenshown to interact in vitro with VP1. An additional viral late protein isthe agnoprotein, or LP1, coded by the leader region of the late 16SmRNA. This small (61 amino acids) protein was found to mediate efficientlocalization of VP1 to the nuclear region and to facilitate release ofmature virus from the infected cells.

It should be appreciated that any mutated viral capsid protein,preferably, VP1 molecule, which posses enhancing effect on chaperonslevels, enhancement of cell survival mechanisms and rescuing cells fromapoptosis or oxidative stress, is encompassed by the present invention.

The viral capsid protein mutants or derivatives thereof that may be usedby the compositions and method of the invention may have at least onemutation selected from the group consisting of point mutations,missense, nonsense, insertions, deletions or rearrangement. It should beappreciated that the terms “insertions” and “deletions” as used hereinis meant any addition or deletions, respectively, of amino acid residuesto the viral capsid proteins of the invention, between 1 to 100 aminoacid residues, preferably, between 1 to 50 amino acid residues and mostpreferably, between 1 to 20 amino acid residues.

According to one particular embodiment, a mutated SV40 VP1 molecule maybe a VP1 molecule deleted in its carboxy terminal arm. Such particularmutated molecule may be the mutant VP1ΔC. According to a particularlypreferred embodiment, this mutated VP1 molecule comprises the amino acidsequence substantially as denoted by SEQ ID NO. 4.

It should be appreciated that additional mutations, including deletionor insertions, may be, for example, in the surface loops of VP1 whichconnect between the β-strands, are within the scope of the invention.Such mutations are expected to be more potent than wild type VP1 inactivating the chaperon machinery and/or survival pathway, as well asprotecting cells against apoptosis or oxidative stress. It should befurther noted that any other mutations affecting the structure of VP1,or VP1 derivative, allowing it to have a prolonged effect in vivo, arealso within the scope of the present invention.

According to a particular embodiment, the compositions of the inventionmay comprise any combination of wild type VP1, the VP1ΔC mutant and anyother mutant described above.

It should be noted that the viral capsid proteins used in thecomposition and methods of the invention can be natural or synthetic,modified or unmodified, whole or fragments thereof. Fragments can bederived from synthesis as fragments or by digestion or other means ofmodification to create fragments from larger entities.

Preferably, the viral capsid proteins may be recombinantly prepared.Preparation of recombinant capsid proteins involves the use of generalmolecular biology techniques that are well known in the art. Suchtechniques include for example, cloning of the gene encoding a desiredviral capsid protein to a suitable expression vector. Preferredrecombinant SV40 VP1 capsid proteins may be prepared using prokaryotic,preferably, bacterial or eukaryotic expression systems. Most preferredare capsid proteins prepared in eukaryotic expression system, such asthe baculovirusdes system described in Experimental procedures. Itshould be noted that construction of these vectors is also described indetail in Sandalon et al., [Sandalon (1997) ibid.], which is a previouspublication by the inventor and is incorporated herein by reference.

It should be further noted that the capsid proteins and specifically theSV40 VP1, may be used for the composition of the invention asdissociated VP1 pentamers or as virus-like particles (VLP's, asdemonstrated by FIG. 1) comprising VP1 and other SV40 capsid proteins oralternatively, containing VP1 only. Still further, it should beappreciated that the viral capsid proteins may be mixed with any otherviral protein such as the agnoprotein or any other protein facilitatingtheir action. Still further, it should be noted that for preparation ofdissociated pentamers or fragments of said SV40 VP1 protein, a bacterialexpression system may be used. Moreover, it should be appreciated thatthe invention further encompasses the use of any fusion proteincomprising said VP1, or any mutant of VP1 (for example, the VP1ΔC), asan active ingredient in any one the compositions of the invention.

According to a specifically preferred embodiment, the pathologicdisorder to be treated with the composition of the invention, may beassociated with inactivation of cellular proteins participating inquality control processes and therefore may be ameliorated by activationof such proteins.

Newly synthesized proteins have a string-like structure, and the stringsmust be adequately folded sterically so that the resultant proteins worknormally. For this purpose, molecules called “molecular chaperons” existin the endoplasmic reticula to help sterically adequate folding of theprotein strings. However, proteins are sometimes not adequately foldedeven with the molecular chaperon's help. Such unusual proteins areexcluded from the endoplasmic reticula (ER) and decomposed in aprotein-degradation factory called “proteasome.” That is, theendoplasmic reticulum possesses a function to distinguish inadequatelyfolded proteins from folded ones in order to dispose of the unusualproteins. This functional mechanism is referred to as the“quality-control mechanism of proteins in the endoplasmic reticulum.” Ifthis mechanism is broken, cells cannot judge which proteins are adequate“products,” inducing severe, life-threatening damage.

More specifically, in living cells, both newly made and preexistingpolypeptide chains are at constant risk for misfolding and aggregation.In accordance with the wide diversity of misfolded forms, elaboratequality-control strategies have evolved to counter these inevitablemishaps. Recent reports describe the removal of aggregates from thecytosol, reveal mechanisms for protein quality control in theendoplasmic reticulum, and provide new insight into two classes ofmolecular chaperones, the Hsp70 system and the AAA+ (Hsp100) unfoldases.

As indicated above, the endoplasmic reticulum (ER) is responsible forthe structural maturation of the roughly one-quarter of the proteomethat traverses the secretory pathway. The ER employs two distinctmechanisms for responding to the presence of misfolded forms. The firstis an ER-dedicated stress response termed the unfolded protein response(UPR), which acts to remodel the ER so as to increase its foldingcapacity. In yeast, the folding capacity of the ER is monitored by IRE1,a highly conserved transmembrane kinase that contains a lumenal domainresponsible for sensing misfolded forms and cytosolic kinase andribonuclease domains. The accumulation of misfolded proteins in the ERleads to activation of the IRE kinase.

The second, termed ER-associated degradation (ERAD), specificallyrecognizes terminally misfolded proteins and retrotranslocates themacross the ER membrane into the cytosol, where they can be degraded bythe ubiquitin-proteasome degradation machinery. As might be expected bythe diversity of proteins that fold in the ER, recent studies argue thatER-associated degradation (ERAD) encompasses a number of differentsystems, each responsible for the degradation of subsets of proteinsthat share common physical properties. This is perhaps most clearlyshown in yeast, where there are at least two distinct surveillancemechanisms for identifying terminally misfolded ER proteins. The first,designated ERAD-L, inspects for proteins that contain misfolded lumenaldomains. The second, termed ERAD-C, detects misfolded cytosolic domainsof transmembrane proteins. Although both of these pathways ultimatelyconverge on the ubiquitin-proteasome degradation system, they depend ondifferent sets of ER-associated components to detect and delivermisfolded species to the cytosol. In the case of ERAD-C (but notERAD-L), degradation is typically dependent on a specific subset ofcytosolic chaperones including Hsp70 and Hsp40 members.

Recent studies have identified two different ER-localized lectins thatplay a critical role in ERAD. The first is related to the mannosidaseprotein responsible for the trimming of N-glycans in the ER but appearsto have lost its catalytic activity. The second lectin, Yos9p, forms astable complex with misfolded proteins, and loss of Yos9p leads to aprofound and specific defect in degradation of misfolded glycoproteins.

The ubiquitin-proteasome pathway might be the mainstay of removal ofaggregation-prone species. It appears now that ubiquitin modificationmay in fact recruit aggregated species for clearance via an independentmechanism, the “autophagy” pathway. Autophagy involves the recognitionand packaging/engulfment of targeted proteins or organelles intoautophagosome vesicles that become fused with lysosomes, wherein bothvesicles and their contents are broken down. More than 20 so-called Atgcomponents mediate this remarkable process.

It has become increasingly apparent that there are a variety ofconditions in vivo where, even with chaperones and the proteolyticmachinery present in the same compartment as a misfolding protein, thesemechanisms of quality control fail and the misfolded proteins proceed toform aggregates. Moreover, such intracellular aggregates are associatedwith a number of neurodegenerative diseases such as Huntington's,Alzheimer's, Amyotrophic lateral sclerosis and Parkinson's. The natureand fate of protein aggregates in eukaryotic cells has been poorlyunderstood.

Without being bound by any theory, as shown by the following Examples,the viral capsid proteins, particularly, VLPs containing VP1, VP1pentamers or mutated VP1 molecule, preferably, the VP1ΔC mutant,activated proteins participating in quality control processes,specifically, chaperones (as shown by the present invention for HSP/c70,for example), and thereby led to clear amelioration of the pathologiccondition treated. This amelioration of a pathologic condition wasclearly demonstrated by the invention using the AKI (acute kidneyinjury) mice model, which was formally referred to as ATN.

Therefore, according to a specific embodiment, proteins participating inquality control processes may be chaperones. For example, chaperones ofthe following families: HSP-40, HSP-70, CALNEXIN, BiP, HSP-27, HSP-22,HSP-60, HSP-65, HSP-27, HSP-90.

Heat shock proteins (HSPs) are called stress proteins or molecularchaperons that assist cell rescue through the folding of synthesized orstress-denatured proteins. As indicated above, HSPs are also believed tobe causative factors in various autoimmune diseases, whose etiologiesare considered to spring from immune responses against HSPs as a targetmolecule. For instance, elevated antibody titers to HSP60 familyproteins as well as the HSP70 family proteins (HSP70 and HSC70), arereported in cerebrospinal fluid (CSF) of patients with multiplesclerosis (MS) and other neurological diseases, and therefore may playan important role in the pathophysiology of MS through the modificationof immune response and cytoprotective functions of molecular chaperons.

Mutations of the heat shock proteins 27 (HSP27) and HSP22, wereidentified in Charcot-Marie-Tooth disease (CMT), which is one of themost common but heterogeneously hereditary motor and sensory neuropathyand in distal hereditary motor neuropathy (HMN) which is an exclusivelymotor neuropathy but is also a clinically and genetically heterogenousneuropathy.

Acute Respiratory Distress Syndrome (ARDS) (an example for inflammatoryresponse), Legioner disease, GBS (Guillain-Barre Syndrome) and podocyteinjury, are also disorders shown to be associated with disfunction ofchaperones, and are therefore conditions that may be treated by thecompositions and methods of the invention.

According to another embodiment, the composition of the invention may beparticularly applicable and suitable for the treatment of pathologicdisorders such as neurodegenerative disorders or immune-relateddisorders. It should be noted that, as indicated above, the pathologicdisorder to be treated by the composition of the invention is preferablya disorder associated with inactivation of cellular proteins involvedwith quality control processes within the cell.

More specifically, immune disorders related to an imbalance in theTh1-Th2 response and therefore, an immune-related disorder may be forexample, an autoimmune disease, (for example, multiple sclerosis (MS),Type-1 diabetes, lupus, Graves disease and thyroiditis), malignant andnon-malignant proliferative disorders, graft rejection pathology andgraft versus host disease, inflammation and also pathogen relateddisorders (such as toxic shock, incapacitation and death, septic shockand severe sepsis, induced by a pyrogenic exotoxin).

Inflammation includes any inflammatory conditions wherein saidinflammatory conditions may be any one of rheumatoid arthritis, acuterespiratory distress syndrome (ARDS), asthma, rhinitis, idiopathicpulmonary fibrosis, peritonitis, cardiovascular inflammation, myocardialischemia, reperfusion injury, atherosclerosis, sepsis, trauma, diabetestype II, retinopathy, psoriasis, gastrointestinal inflammation,cirrhosis and inflammatory bowel disease.

In general, the compositions as well as the methods of the presentinvention described herein below, may be used in the treatment of anyautoimmune disease such as for example, but not limited to,Eaton-Lambert syndrome, Goodpasture's syndrome, Greave's disease,Guillain-Barr syndrome, autoimmune hemolytic anemia (AIHA), hepatitis,insulin-dependent diabetes mellitus (IDDM), systemic lupus erythematosus(SLE), multiple sclerosis (MS), myasthenia gravis, plexus disorders e.g.acute brachial neuritis, polyglandular deficiency syndrome, primarybiliary cirrhosis, rheumatoid arthritis, scleroderma, thrombocytopenia,thyroiditis e.g. Hashimoto's disease, Sjogren's syndrome, allergicpurpura, psoriasis, mixed connective tissue disease, polymyositis,dermatomyositis, vasculitis, polyarteritis nodosa, polymyalgiarheumatica, Wegener's granulomatosis, Reiter's syndrome, Behget'ssyndrome, ankylosing spondylitis, pemphigus, bullous pemphigoid,dermatitis herpetiformis, insulin dependent diabetes, inflammatory boweldisease, ulcerative colitis and Crohn's disease.

As used herein to describe the present invention, the terms “malignantproliferative disorder”, “cancer”, “tumor” and “malignancy” all relateequivalently to a hyperplasia of a tissue or organ. If the tissue is apart of the lymphatic or immune systems, malignant cells may includenon-solid tumors of circulating cells. Malignancies of other tissues ororgans may produce solid tumors. In general, the composition as well asthe methods of the present invention described below, may be used in thetreatment of non-solid and solid tumors, for example, carcinoma,melanoma, leukemia, and lymphoma.

Therefore, according to a preferred embodiment, the SV40 capsid proteinsor any composition comprising the same according to the invention, canbe used for the treatment or inhibition of non-solid cancers, e.g.hematopoietic malignancies such as all types of leukemia, e.g. acutelymphocytic leukemia (ALL), acute myelogenous leukemia (AML), chroniclymphocytic leukemia (CLL), chronic myelogenous leukemia (CML),myelodysplastic syndrome (MDS), mast cell leukemia, hairy cell leukemia,Hodgkin's disease, non-Hodgkin's lymphomas, Burkitt's lymphoma andmultiple myeloma, as well as for the treatment or inhibition of solidtumors such as tumors in lip and oral cavity, pharynx, larynx, paranasalsinuses, major salivary glands, thyroid gland, esophagus, stomach, smallintestine, colon, colorectum, anal canal, liver, gallbladder,extraliepatic bile ducts, ampulla of Vater, exocrine pancreas, lung,pleural mesothelioma, bone, soft tissue sarcoma, carcinoma and malignantmelanoma of the skin, breast, vulva, vagina, cervix uteri, corpus uteri,ovary, fallopian tube, gestational trophoblastic tumors, penis,prostate, testis, kidney, renal pelvis, ureter, urinary bladder,urethra, carcinoma of the eyelid, carcinoma of the conjunctiva,malignant melanoma of the conjunctiva, malignant melanoma of the uvea,retinoblastoma, carcinoma of the lacrimal gland, sarcoma of the orbit,brain, spinal cord, vascular system, hemangiosarcoma and Kaposi'ssarcoma.

In yet another specific embodiment, the compositions of the invention aswell as the methods of the invention described herein after, may be usedfor the treatment of a neurodegenerative disorder.

A “neurological disorder” is a disease or disorder characterized by anabnormality or malfunction of neuronal cells or neuronal support cells.The disorder can affect the central and/or peripheral nervous system.Exemplary neurological diseases include neuropathies, skeletal muscleatrophy and neurodegenerative diseases.

“Neurodegenerative disorders” are complex and pernicious diseases, theironset is insidious, followed by progressive deterioration. Clinicalmanifestations are determined by the location and seriousness of thedisorder. Although the causes may differ, patients withneurodegenerative disorders are likely to show localized to generalizedatrophy of brain cells, leading to compromises in both mental andphysical function. Exemplary neurodegenerative diseases include:Alzheimer's disease, Parkinson's disease, ALS (Amyotrophic LateralSclerosis), Huntington's disease, taupathies such as Pick's disease,fronto temporal dementia, cortico-basal degeneration and progressivesupranuclear palsy and Spongiform encephalopathies such as Scrapie, madcow disease and Bovine spongiform encephalopathy, Creutzfeldt-Jakobdisease, Fatal Familial Insomnia, Gerstmann-Straussler-Scheinkersyndrome and Kuru.

As indicated above, the pathologic disorder treated by the compositionof the invention may be a disorder induced by a pathogenic agent.Pathogenic agents include prokaryotic microorganisms, lower eukaryoticmicroorganisms, complex eukaryotic organisms, viruses, fungi, prions,parasites, yeasts, toxins and venoms.

A prokaryotic microorganism includes bacteria such as Gram positive,Gram negative and Gram variable bacteria and intracellular bacteria.Examples of bacteria contemplated herein include the species of thegenera Treponema sp., Borrelia sp., Neisseria sp., Legionella sp.,Bordetella sp., Escherichia sp., Salmonella sp., Shigella sp.,Klebsiella sp., Yersinia sp., Vibrio sp., Hemophilus sp., Rickettsiasp., Chlamydia sp., Mycoplasma sp., Staphylococcus sp., Streptococcussp., Bacillus sp., Clostridium sp., Corynebacterium sp.,Proprionibacterium sp., Mycobacterium sp., Ureaplasma sp. and Listeriasp.

Particular species include Treponema pallidum, Borrelia burgdorferi,Neisseria gonorrhea, Neisseria meningitidis, Legionella pneumophila,Bordetella pertussis, Escherichia coli, Salmonella typhi, Salmonellatyphimurium, Shigella dysenteriae, Klebsiella pneumoniae, Yersiniapestis, Vibrio cholerae, Hemophilus influenzae, Rickettsia rickettsii,Chlamydia trachomatis, Mycoplasma pneumoniae, Staphylococcus aureus,Streptococcus pneumoniae, Streptococcus pyogenes, Bacillus anthracis,Clostridium botulinum, Clostridium tetani, Clostridium perfringens,Corynebacterium diphtheriae, Proprionibacterium acnes, Mycobacteriumtuberculosis, Mycobacterium leprae and Listeria monocytogenes.

A lower eukaryotic organism includes a yeast or fungus such as but notlimited to Pneumocystis carinii, Candida albicans, Aspergillus,Histoplasma capsulatum, Blastomyces dermatitidis, Cryptococcusneoformans, Trichophyton and Microsporum.

A complex eukaryotic organism includes worms, insects, arachnids,nematodes, aemobe, Entamoeba histolytica, Giardia lamblia, Trichomonasvaginalis, Trypanosoma brucei gambiense, Trypanosoma cruzi, Balantidiumcoli, Toxoplasma gondii, Cryptosporidium or Leishmania.

The term “viruses” is used in its broadest sense to include viruses ofthe families adenoviruses, Papilloma and polyomaviruses, herpesviruses:simplex, varicella-zoster, Epstein-Barr, CMV, pox viruses: smallpox,vaccinia, hepatitis B, rhinoviruses, hepatitis A, poliovirus, rubellavirus, hepatitis C, arboviruses, rabies virus, influenza viruses A andB, measles virus, mumps virus, HIV, HTLV I and II.

The term “fungi” includes for example, fungi that cause diseases such asringworm, histoplasmosis, blastomycosis, aspergillosis, cryptococcosis,sporotrichosis, coccidioidomycosis, paracoccidio-idoinycosis, andcandidiasis.

The term parasite includes, but not limited to, infections caused bysomatic tapeworms, blood flukes, tissue roundworms, ameba, andPlasmodium, Trypanosoma, Leishmania, and Toxoplasma species.

According to a particular and preferred embodiment, the compositions ofthe invention is intended for the treatment of pathologic disorderinduced by pathogenic agent. Such disorders may lead to sepsis which mayresults in any one of ARF (acute renal failure) condition or diseasesuch as AKI, ARDS (acute respiratory distress syndrome) and multiplesystem organ failure.

According to a particular and preferred embodiment, and as clearlydemonstrated by Examples 3-7, the composition of the invention isparticularly suitable for the treatment of ARF as demonstrated using theAKI model, which as indicated above, is a disorder which can be relievedby activation or addition of chaperones.

Acute kidney injury (AKI), formerly referred to as “acute tubularnecrosis” encompasses a host of physiopathological mechanisms. Directtubular toxicity, endothelial dysfunction with altered renalmicrocirculation, tubular hypoxic injury, damage from reactive oxygenspecies, impaired glomerular hemodynamics and localized or systemicinflammation, all interact to form renal dysfunction, characterized byacute reduction of glomerular filtration rate (GFR) and defectivetubular function. Progression of chronic kidney disease (CKD) alsoreflects a host of physiopathological mechanisms, initiated following aprimary renal structural damage. Three major instigating factors,glomerular hyperfiltration, proteinuria and renal parenchymal hypoxialead to microvascular and nephron atrophy and to interstitial fibrosisvia multiple compound and interacting mechanisms. Both AKI and theprogression of CKD to end-stage renal failure lead to substantialmorbidity and to markedly increased mortality, and are associated withlarge expenses, required for their clinical management.

Though the physiologic mechanisms associated with these disorders areonly partially understood, renal parenchymal apoptotic or non-apoptoticcell death evidently plays a central role, and measures to attenuatethese processes are a major goal in therapeutic interventions. One suchpotential intervention is the induction of cellular adaptive mechanisms,such as the upregulation of stress response genes, including heat shockproteins (HSP). These highly conserved molecules along the evolutiontree act as chaperons for injured proteins throughout their proteasomaldegradation [Aufricht, C. Pediatr. Nephrol. 20: 707-713 (2005)]. HSPsare considered cell protective and their induction by transient hypoxiaor heat stress were shown to attenuate apoptotic damage and AKI both invitro and in vivo [Lu, C. Y. et al. Curr. Opin. Nephrol. Hypertens.16:83-89 (2007)].

It should be noted that all disorders indicated herein as disorders thatmay be treated by the compositions of the invention may also be treatedby the methods of the invention described herein after.

According to another particular embodiment, the composition of theinvention is provided for use in the treatment of ARF, and particularly,AKI.

As shown by Examples 2 and 7, VLP's comprising SV40 VP1 protect kidneycells from apoptosis, and may also protect cells from oxidative stress.Therefore, it should be noted that the compositions of the invention,may be further used in vitro or ex vivo for rescuing cells in a subjectin need thereof from pathologic processes involving oxidative stress andapoptosis.

The term apoptosis, or programmed cell death, is a normal component ofthe development and health of multicellular organisms. Cells die inresponse to a variety of stimuli and during apoptosis, they do so in acontrolled, regulated fashion. This makes apoptosis distinct fromanother form of cell death called necrosis in which uncontrolled celldeath leads to lysis of cells, inflammatory responses and, potentially,to serious health problems. However, as indicated herein before,uncontrolled apoptotic process in some cases, may lead to pathologicdisorders, for example, AKI, and thus, should be prevented. Theinvention therefore provides methods and compositions for attenuatingharmful apoptotic process an induce cell survival pathways.

As indicated above, the composition of the invention may be applicablefor conditions involving oxidative stress. Oxidative stress is caused byan imbalance between the production of reactive oxygen and a biologicalsystem's ability to readily detoxify the reactive intermediates oreasily repair the resulting damage. All forms of life maintain areducing environment within their cells. The cellular redox environmentis preserved by enzymes that maintain the reduced state through aconstant input of metabolic energy. Disturbances in this normal redoxstate can cause toxic effects through the production of peroxides andfree radicals that damage all components of the cell, includingproteins, lipids, and DNA. Wide variety of diseases have evidence ofexcess generation of free radicals, oxidative stress and inadequateantioxidant activity. Some examples are neurodegenerative diseases (seebelow), heart disease, HIV disease, chronic fatigue syndrome, hepatitis,cancer, autoimmune diseases, etc.

As indicated by the Examples, unpublished data of the inventors indicatethat SV40 VP1 rescue cells from oxidative stress and apoptosis, probablyby induction of a cell survival mechanism, preferably, the PI3K-PKB/Aktsurvival pathway. Therefore, according to one embodiment, thecomposition of the present invention may be used for inductions of cellsurvival pathway, preferably, the PI3K-PKB/Akt survival pathway, incells of a subject in need thereof.

It should be noted that PKB/Akt is a key player in many cellularsurvival mechanisms. It has been implicated, in particular, in potentialneuroprotection in neurodegenerative diseases [Schmeer, C. et al. RestorNeurol Neurosci 24:79-95 (2006)], including parkinson's disease [Fallon,L. et al. Nat. Cell. Biol. 8:834-842 (2006)], Alzheimer's disease [Cole,G. M. et al. Exp. Gerontol. 42:10-21 (2007)], and in neural survivalafter stroke and brain injury [Zhang, X. J. Cereb. Blood Flow Metab.26:915-926 (2006)]. Additional examples are protection againstcardiohypeprtrophy [Patten, R. D. and Karas, R. H. Trends Cardiovasc.Med. 16:69-75 (2006)] and kidney diseases [Mitch, W. E. J. Ren. Nutr.17:66-69 (2007)].

Still further, the present invention provides a pharmaceuticalcomposition for enhancing the ameliorating effect of chaperones onpathologic disorders.

It should be appreciated that the pharmaceutical compositions of theinvention generally comprise a buffering agent, an agent which adjuststhe osmolarity thereof, and optionally, one or more pharmaceuticallyacceptable carriers, excipients and/or additives as known in the art.Supplementary active ingredients can also be incorporated into thecompositions. The carrier can be solvent or dispersion mediumcontaining, for example, water, ethanol, polyol (for example, glycerol,propylene glycol, and liquid polyethylene glycol, and the like),suitable mixtures thereof, and vegetable oils. The proper fluidity canbe maintained, for example, by the use of a coating, such as lecithin,by the maintenance of the required particle size in the case ofdispersion and by the use of surfactants.

As used herein “pharmaceutically acceptable carrier” includes any andall solvents, dispersion media, coatings, antibacterial and antifungalagents and the like. The use of such media and agents for pharmaceuticalactive substances is well known in the art. Except as any conventionalmedia or agent is incompatible with the active ingredient, its use inthe therapeutic composition is contemplated.

In a second aspect, the present invention relates to a method for thetreatment of a pathologic disorder in a subject in need thereof. Thismethod comprises the step of administering to the treated subject atherapeutically effective amount of at least one viral capsid protein orany peptide, mutant or fragment thereof, or of VLPs comprising at leastone capsid protein or any peptide, mutant or fragment thereof, or of acomposition comprising the same.

The method of the invention may be applicable for treating a subjectsuffering from a pathologic disorder such as a neurodegenerativedisorder or an autoimmune-related disorder. As used herein, the term“disorder” refers to a condition in which there is a disturbance ofnormal functioning. A “disease” is any abnormal condition of the body ormind that causes discomfort, dysfunction, or distress to the personaffected or those in contact with the person. Sometimes the term is usedbroadly to include injuries, disabilities, syndromes, symptoms, deviantbehaviors, and atypical variations of structure and function, while inother contexts these may be considered distinguishable categories. Itshould be noted that the terms “disease”, “disorder”, “condition” and“illness”, are equally used herein.

The terms “treat, treating, treatment” as used herein and in the claimsmean ameliorating one or more clinical indicia of disease activity in apatient having a pathologic disorder.

“Treatment” refers to therapeutic treatment. Those in need of treatmentare mammalian subjects suffering from any pathologic disorder or anautoimmune-related disorder. By “patient” or “subject in need” is meantany mammal for which administration of the viral capsid proteins, or anypharmaceutical composition of the invention is desired, in order toprevent, overcome or slow down such infliction.

To provide a “preventive treatment” or “prophylactic treatment” isacting in a protective manner, to defend against or prevent something,especially a condition or disease.

According to one embodiment, the viral capsid protein may be any one ofthe capsid proteins of papillomaviruses or polyomaviruses, or anyfragment, peptide, mutant, any mixtures and combinations thereof.Preferably, the viral capsid protein may be at least one of SV40 VP1,VP2, VP3, and any peptide, fragment, mutant, mixtures and combinationsthereof. Most preferably, the viral capsid protein may be SV40 VP1, anypeptide, mutant or fragment thereof or any VLPs comprising at SV40 VP1or any peptide, mutant or fragment thereof. Particular embodimentrelates to the SV40 VP1 protein comprising the amino acid sequence asdenoted by SEQ ID NO: 1.

According to another specific embodiment, a mutated SV40 VP1 moleculemay be the VP1ΔC mutant. Such mutated molecule may preferably comprisethe amino acid sequence as substantially denoted by SEQ ID NO. 4.

Preferably, the pathologic disorder treated by the method of theinvention are conditions caused by inactivation of cellular proteinsparticipating in quality control processes or in inflammatory processesand therefore, may be ameliorated by activation of such cellularproteins.

According to a specifically preferred embodiment, the proteinsparticipating in quality control processes may be chaperones. As clearlydemonstrated by the invention, administration of VLP's containing VP1,or VP1 pentamers composed of the VP1ΔC mutated molecule, resulted inelevation in chaperon levels, particularly of chaperone HSP/c70. Thiselevation was correlated with the beneficial effect of SV40 VP1, on ARFconditions such as AKI, as clearly demonstrated by survival of treatedanimals, reduced blood urea and creatinine levels.

According to another embodiment, the method of the invention is intendedfor the treatment of a pathologic disorder such as a neurodegenerativedisorder or an immune-related disorder.

According to a specific embodiment, the method of the invention issuitable for the treatment of an immune-related disorder such as anautoimmune disease, malignant and non-malignant proliferative disorders,graft rejection pathology and graft versus host disease, and disordersinduced by a pathogenic agent.

In yet another specifically preferred embodiment, the method of theinvention is specifically suitable for the treatment of pathologicdisorder induced by pathogenic agent. Such disorders may lead to sepsiswhich may results in any one of ARF (acute renal failure), ARDS (acuterespiratory distress syndrome) and multiple system organ failure.

Therefore, according to a specifically preferred embodiment, said ARFdisease may be AKI.

As indicated herein before, acute Kidney Injury (AKI) is characterizedpathologically by varying degrees of tubular cell damage. Renal celldeath may result from prolonged renal ischemia, nephrotoxins, or sepsis.AKI results clinically in a rapid (hours to days) decline in theglomerular filtration rate (GFR) and tubular function that lead toretention of uremic toxins, acid-base disturbance, fluid imbalance anddysfunction of virtually all other systems.

The terms “effective amount” or “sufficient amount” as used by themethods of the invention, mean an amount necessary to achieve a selectedresult. The “effective treatment amount” is determined by the severityof the disease in conjunction with the preventive or therapeuticobjectives, the route of administration and the patient's generalcondition (age, sex, weight and other considerations known to theattending physician).

Therapeutic formulations may be administered in any conventional dosageformulation. According to a particular and specific embodiment, theinvention uses viral capsid proteins for the treatment of ARF, asdemonstrated by the AKI model. As shown by the examples, the SV40 VP1protein was used as VLPs, which were injected to the diseased animals.Appropriate dose may range between 0.01 to 100 mg/kg of weight,preferably, about 0.1 to 50 mg/kg, most preferably 0.1 to 20 mg/kg.Formulations typically comprise at least one active ingredient, asdefined above, together with one or more acceptable carriers thereof.

As shown by the Examples, also mutated SV40 VP1 molecule, the VP1ΔC, maybe used by the compositions and the methods of the invention, althoughhigher amount may be required for achieving the desired beneficialeffect on treated cells.

It should be noted that formulations used by the compositions andmethods of the invention include those suitable for oral, rectal, nasal,or parenteral (including subcutaneous, intramuscular, intravenous andintradermal) administration. The formulations may conveniently bepresented in unit dosage form and may be prepared by any methods wellknown in the art of pharmacy. The nature, availability and sources, andthe administration of all such compounds including the effective amountsnecessary to produce desirable effects in a subject are well known inthe art and need not be further described herein.

In yet another embodiment, the administering step according to themethod of the invention, comprises oral, intravenous, intramuscular,subcutaneous, intraperitonea, perenteral, transdermal, intravaginal,intranasal, mucosal, sublingual, topical, rectal or subcutaneousadministration, or any combination thereof.

As indicated above, pharmaceutical compositions for use in accordancewith the present invention may be formulated in conventional mannerusing one or more physiologically acceptable carriers comprisingexcipients and auxiliaries, which facilitate processing of the activeingredients into preparations which, can be used pharmaceutically.Proper formulation is dependent upon the route of administration chosen.

For injection, the active ingredients of the invention may be formulatedin aqueous solutions, preferably in physiologically compatible bufferssuch as Hank's solution, Ringer's solution, or physiological saltbuffer. For transmucosal administration, penetrants appropriate to thebarrier to be permeated are used in the formulation. Such penetrants aregenerally known in the art.

Pharmaceutical compositions for topical administration may includetransdermal patches, ointments, lotions, creams, gels, drops,suppositories, sprays, liquids and powders. Conventional pharmaceuticalcarriers, aqueous, powder or oily bases, thickeners and the like may benecessary or desirable.

For oral administration, the compounds can be formulated readily bycombining the active compounds with pharmaceutically acceptable carrierswell known in the art. Such carriers enable the compounds of theinvention to be formulated as tablets, pills, dragees, capsules,liquids, gels, syrups, slurries, suspensions, and the like, for oralingestion by a patient.

Pharmacological preparations for oral use can be made using a solidexcipient, optionally grinding the resulting mixture, and processing themixture of granules, after adding suitable auxiliaries if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol, cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarbomethylcellulose; and/or physiologically acceptable polymers such aspolyvinylpyrrolidone (PVP). Thickeners, flavoring agents, diluents,emulsifiers, dispersing aids or binders may be desirable.

For administration by nasal inhalation, the active ingredient for useaccording to the present invention, which is the viral capsid proteins,or preferably, VLP's comprising SV40 VP1, or mutated VP1 molecule, mayconveniently delivered in the form of an aerosol spray presentation froma pressurized pack or a nebulizer with the use of a suitable propellant,e.g., dichlorodifluoromethane, trichlorofluoromethane,dichloro-tetrafluoroethane or carbon dioxide. In the case of apressurized aerosol, the dosage unit may be determined by providing avalve to deliver a metered amount. Capsules and cartridges of, e.g.,gelatin for use in a dispenser may be formulated containing a powder mixof the compound and a suitable powder base such as lactose or starch.

The preparations described herein may be formulated for parenteraladministration, e.g., by bolus injection or continuous infusion.Formulations for injection may be presented in unit dosage form, e.g.,in ampoules or in multidose containers with optionally, an addedpreservative. The compositions may be suspensions, solutions oremulsions in oily or aqueous vehicles, and may contain formulatoryagents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical compositions for parenteral administration includeaqueous solutions of the active preparation in water-soluble form.Additionally, suspensions of the active ingredients may be prepared asappropriate oily or water based injection suspensions. Suitablelipophilic solvents or vehicles include fatty oils such as sesame oil,or synthetic fatty acids esters such as ethyl oleate, triglycerides orliposomes. Aqueous injection suspensions may contain substances, whichincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension may alsocontain suitable stabilizers or agents, which increase the solubility ofthe active ingredients to allow for the preparation of highlyconcentrated solutions.

Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile, pyrogen-free waterbased solution, before use.

The pharmaceutical composition of the present invention may also beformulated in rectal compositions such as suppositories or retentionenemas, using, e.g., conventional suppository bases such as cocoa butteror other glycerides.

Thus, the pharmaceutical compositions of the present invention include,but are not limited to, solutions, emulsions, and liposome-containingformulations. These compositions may be generated from a variety ofcomponents that include, but are not limited to, preformed liquids,self-emulsifying solids and self-emulsifying semisolids.

Pharmaceutical compositions suitable for use in context of the presentinvention include compositions wherein the active ingredients arecontained in an amount effective to achieve the intended purpose. Morespecifically, a therapeutically effective amount means an amount ofactive ingredients effective to prevent, alleviate or amelioratesymptoms of disease or prolong the survival of the subject beingtreated.

As indicated above, determination of a therapeutically effective amountis well within the capability of those-skilled in the art.

For any pharmaceutical composition used by the treatment method of theinvention, the therapeutically effective amount or dose can be estimatedinitially from in vitro assays. For example, a dose can be formulated inanimal models and such information can be used to more accuratelydetermine useful doses in humans.

Toxicity and therapeutic efficacy of the active ingredients describedherein can be determined by standard pharmaceutical procedures in vitro,in cell cultures or experimental animals. The data obtained from invitro cell culture assays and animal studies can be used in formulatinga range of dosage for use in human. The dosage may vary depending uponthe dosage form employed and the route of administration utilized. Theexact formulation, route of administration and dosage can be chosen bythe individual physician in view of the patient's condition.

Depending on the severity and responsiveness of the condition to betreated, dosing can be of a single or a plurality of administrations,with course of treatment lasting from several days to several weeks oruntil cure is effected or diminution of the disease state or symptoms isachieved.

The amount of the pharmaceutical composition to be administered will ofcourse be dependent on the subject being treated, the severity of theaffliction, the manner of administration, the judgment of theprescribing physician, etc.

Although the method of the invention is particularly intended for thetreatment of pathologic disorders in humans, other mammals are included.By way of non-limiting examples, mammalian subjects include monkeys,equines, cattle, canines, felines, mice, rats and pigs.

In a third aspect, the invention relates to a method for enhancing theameliorating effect of cellular proteins participating on qualitycontrol processes, on a pathologic disorder. This method comprises thesteps of contacting, ex vivo or in vitro, cells obtained from a subjectsuffering of a pathologic disorder with an effective amount of at leastone viral capsid protein or any peptides or fragments thereof.

The invention further provides a method of enhancing the amelioratingeffect of cellular proteins participating on quality control processeson a pathologic disorder in a subject in need thereof. This methodcomprises the step of administering to said subject, a therapeuticallyeffective amount of at least one viral capsid protein or any peptides orfragments thereof, or of VLP's comprising the same, or of a compositioncomprising the same.

According to a specifically preferred embodiment, cellular proteinsparticipating on quality control processes may be chaperones,particularly, HSP/c70.

The invention further provides a method for rescuing cells fromoxidative stress or apoptosis, comprising the step of contacting cellsundergoing oxidative stress or apoptotic process, with an effectiveamount of at least one viral capsid protein or any peptide, mutant orfragment thereof or of VLP's comprising said viral capsid protein or anypeptide, mutant or fragment thereof, or of a composition comprising thesame.

According to one particular embodiment, these cells are cells of asubject suffering of a pathologic disorder involving apoptotic processor oxidative stress.

The invention further provides methods for enhancing cell survivalpathway, preferably, the PI3K-PKB/Akt survival pathway. The method ofthe invention comprises the step of contacting cells with effectiveamount of at least one viral capsid protein or any peptide, mutant orfragment thereof or of VLP's comprising the same. According to oneparticular embodiment, induction of cell survival pathway, by themethods and compositions of the invention may be performed in vitro orin vivo in cell culture, or alternatively, in vivo, in a subject in needthereof.

According to one embodiment, the viral capsid protein provided by any ofthe methods of the invention may be any capsid protein ofpapillomaviruses or polyomaviruses, or any fragment, peptides andmixtures and combinations thereof. Preferably, the viral capsid proteinmay be at least one of SV40 VP1, VP2, VP3, and any peptide, fragment,mutant, any mixtures and combinations thereof. Most preferably, theviral capsid protein may be SV40 VP1 or any peptide, mutant or fragmentthereof, or VLP's comprising SV40 VP1. Specifically, the SV40 VP1protein may comprise the amino acid sequence as denoted by SEQ ID NO: 1.

In yet another embodiment, the mutated VP1 molecule used by the methodof the invention may be the VP1ΔC mutant, which preferably comprises theamino acid sequence as denoted by SEQ ID NO. 4.

Preferably, the method of the invention enhances the ameliorating effectof cellular proteins participating in quality control processes andtherefore may be particularly used in pathologic disorder caused byinactivation of such proteins, preferably, chaperones.

According to another embodiment, the method of the invention enhancesthe ameliorating effect of cellular proteins, preferably, chaperones, ona pathologic disorder such as a neurodegenerative disorder or animmune-related disorder.

According to a specific embodiment, an immune-related disorder may be anautoimmune disease, malignant and non-malignant proliferative disorders,graft rejection pathology and graft versus host disease, and disordersinduced by a pathogenic agent.

In yet another specifically preferred embodiment, the pathologicdisorder induced by pathogenic agent may lead to sepsis which mayresults in any one of ARF diseases, ARDS (acute respiratory distresssyndrome) and multiple system organ failure.

Therefore, according to a specifically preferred embodiment, such ARFdisorder may be AKI.

In a fourth aspect, the invention relates to the use of at least oneviral capsid protein or any peptide, mutant or fragment thereof in thepreparation of a pharmaceutical composition for the treatment of apathologic disorder.

According to one embodiment, the viral capsid protein used by theinvention may be any capsid protein of papillomaviruses orpolyomaviruses, or any fragment, mutant, peptide, mixtures andcombinations thereof. Preferably, the viral capsid protein may be atleast one of SV40 VP1, VP2, VP3, and any peptide, fragment, mutant,mixture and combinations thereof. Most preferably, the viral capsidprotein may be SV40 VP1 or any peptide, mutant or fragment thereof.

Preferably, the use according to the invention is for the preparation ofa composition which enhances the ameliorating effect of cellularproteins participating in quality control processes and therefore may beparticularly used for the treatment of a pathologic disorder caused byinactivation of such proteins, preferably, of chaperones.

According to another embodiment, the use of viral capsid proteinsaccording to the invention is for preparation of a composition for thetreatment of a pathologic disorder such as a neurodegenerative disorderor an immune-related disorder.

According to a specific embodiment, an immune-related disorder may be anautoimmune disease, malignant and non-malignant proliferative disorders,graft rejection pathology and graft versus host disease, and disordersinduced by a pathogenic agent.

A pathologic disorder induced by pathogenic agent may lead to sepsiswhich may results in any one of ARF disorder, ARDS (acute respiratorydistress syndrome) and multiple system organ failure.

According to a specifically preferred embodiment, such ARF disorder maybe AKI.

The invention further provides the use of at least one viral capsidprotein or any peptide, mutant or fragment thereof, or of VLP'scomprising said capsid protein or any peptide, mutant or fragmentthereof, in the preparation of a pharmaceutical composition for rescuingcells from oxidative stress or apoptosis.

According to one specific embodiment, the mutated VP1ΔC molecule may beused by the invention.

Disclosed and described, it is to be understood that this invention isnot limited to the particular examples, methods steps, and compositionsdisclosed herein as such methods steps and compositions may varysomewhat. It is also to be understood that the terminology used hereinis used for the purpose of describing particular embodiments only andnot intended to be limiting since the scope of the present inventionwill be limited only by the appended claims and equivalents thereof.

It must be noted that, as used in this specification and the appendedclaims, the singular forms “a”, “an” and “the” include plural referentsunless the content clearly dictates otherwise.

Throughout this specification and the Examples and claims which follow,unless the context requires otherwise, the word “comprise”, andvariations such as “comprises” and “comprising”, will be understood toimply the inclusion of a stated integer or step or group of integers orsteps but not the exclusion of any other integer or step or group ofintegers or steps.

The following examples are representative of techniques employed by theinventors in carrying out aspects of the present invention. It should beappreciated that while these techniques are exemplary of preferredembodiments for the practice of the invention, those of skill in theart, in light of the present disclosure, will recognize that numerousmodifications can be made without departing from the spirit and intendedscope of the invention.

EXAMPLES Experimental Procedures AKI (Acute Kidney Injury) Model Animals

AKI or as formally referred as ATN (acute tubular necrosis), are inducedby the nephrotoxic agent HgCl₂, at a dose of 6.5 mg/kg. Injected micedeveloped AKI as seen by their general appearance and as measured byurea level in the blood. Blood urea levels in normal healthy animalsusually range about <50 mg/dL. Blood urea significantly rises in AKIanimals in the first 3-4 days. Most of the animals treated with thisHgCl₂ dose die within 4 days. Those that survive start showing reductionin blood urea starting from day 5 and are likely to recover.

Production of VLPs

Recombinant baculovirus expressing VP1 from the polyhedrin promoter[Sandalon (1997) ibid.; Sandalon, Z. and Oppenheim, A. In SV40protocols, L. Raptis, ed. (Totowa, N.J., Humana Press Inc.) (2001)] werepropagated in Sf9 cells. Briefly, high titer virus stocks (>10⁹ pfu/ml)were used to infect logarithmic cultures of Sf9 cells, at moi 10, forVP1 production. Seventy two hours post infection the cells wereharvested by centrifugation and nuclear extracts were prepared byprocedures adapted from Schreiber et al. [Schreiber et al., NucleicAcids Research 17: 6419 (1989)], or alternatively, were harvested after5-6 days from the medium, following lysis of the infected Sf9 cells.Nuclear extracts were stored in aliquots at −80° C. Such nuclearextracts contain spontaneously assembled SV40 virus-like particles(VLPs). In order to remove macromolecules, mostly RNA, that may betrapped within the VLPs, they were disassembled and reassembled in 3steps as follows: In step A, 5 μl Sf9 nuclear extract were treated with1 μl of 150 mM DTT and 10 μg RNase in a final volume of 10 μl at 37° C.for 20 minutes. In step B reassembly mix (containing 10 mM ATP, 20 mMHepes-KOH buffer at pH 7.9, 80 mM KCl, 40 mM NH₄Cl, 10 mM MgCl₂, 16%Glycerol and 0.08% NP-40, in a final volume of 10 μl) was added to thetreated nuclear extracts (step A), and the reassembly reaction wasincubated at 37° C. for 1 hour. For step C the packaging reaction waskept on ice overnight after the addition of 10 μl stabilization buffer(150 mM sodium acetate buffer pH 5.2, 3 mM CaCl₂, 120 mM KCl and 40 mMNH₄Cl, keeping the salt concentration at 160 mM), bringing the totalvolume to 30 μl.

The reaction mixture was treated with chloroform (5 μl), the mixture wasvortexed, separated by centrifugation and partially purified VLPsrecovered in the aqueous layer. VLPs were further purified andconcentrated ˜300 fold by stirred-ultrafiltration (to a finalconcentration 5-10 mg/ml) under Argon using XM300 membrane (Millipore).The concentrate was re-suspended in 0.5 M NaCl saline (half the originalvolume) and re-filtered 3 additional times. The purified VLPs arealiquoted and stored at −20° C. until use. Just before use the VLPs arediluted 3 fold with H₂O (to a final NaCl concentration of 166 mM) andfurther diluted with saline to the desired VP1 concentration.

EM of Purified VLPs

Transmission electron microscopy pictures of VLPs were performed asfollows:

Samples were adsorbed onto formvar-carbon-coated copper grids andstained with 1% sodium phosphotungstate, pH 7.0. The samples were viewedin a Philips CM-12 electron microscope, using a voltage of 100 kV, andphotographed at a magnification of 66,000×.

Purified VLPs are demonstrated by FIG. 1. As shown by the figure, inboth methods of harvesting (nuclear or medium), VLP's appear as isolatednanoparticles of uniform shape and size (FIG. 1A) similar to wild typeSV40 (FIG. 1B). Gel electrophoresis indicated that they contain ˜95% VP1(FIG. 1C). No DNA was detected in the particles.

Example 1 Induction/Activation of Chaperons (Hsp/c70) by SV40 LVP's

As indicated in the background of the invention, chaperons weredemonstrated as ameliorating clinical conditions such as acute tubularnecrosis (ATN) or as presently defined as, AKI, and SV40 was shown toactivate the expression of chaperone Hsp70. The inventors have thusexamined whether SV40 capsid proteins, without any genetic material,affect or may modulate the levels of chaperone proteins such as Hsp70for example. Therefore, the induction of chaperon biosynthesis by SV40capsid proteins was next evaluated. SV40 capsid proteins as virus likeparticles (VLP's) containing VP1 only and as dissociated VP1 pentamers,were added to CV1 cells, derived from African Green monkey kidney (SV40VLP's, 50 ng and 500 ng per 10⁶ cells). At several time points afteraddition of the capsid proteins, total cellular proteins were harvestedand equal amounts were analyzed by polyacrylamide gel electrophoresisand Western blotting using antibodies specific to mammalian HSP/c70chaperone. Untreated mice served as controls. FIG. 2A shows Western blotindicating levels of Hsp/c70 protein. The enhancement of HSP/c70expression by SV40 VLPs was further analyzed. As shown by FIG. 2B, CV1cells were treated with 50 ng VLPs, immunostained with anti-Hsp/c 70antibody and viewed under confocal microscopy. As clearly demonstratedby the figure, Monkey CV-1 cells respond to VLP's infection byupregulating Hsp/c70. The Western analysis showed significant increasein protein level at 9 hours PI, at both concentrations of VP1 (50 and500 ng). Confocal microscopy (FIG. 2B) showed a slight increase in thelevel of Hsp/c70 at 6 hours. Hsp/c70 continued to accumulate in thecells and is translocated towards the nucleus by 9 hours PI. At 48 hoursHsp/c70 is no longer seen. Similar results were obtained using themutated VP1ΔC molecule (data not shown). The inventors thereforeconclude that there is an increase in Hsp/c70, starting at 6-9 hours PI.

Example 2 SV40 VLPs Protect Cultured Kidney Cells Against Apoptosis

Accumulating evidence suggest that chaperones protect against renalfailure [Lu, C. Y. et al. Curr. OP. in Nephrol. Hypertens. 16:83-89(2007); Riordan, M. et al. Nat. Clin. Pract. Nephrol. 2:149-156 (2006);Kelly, K. J. Contrib. Nephrol. 148:86-106 (2005)]. Therefore, theinventors have next examined whether the induction of hsp/c70 by SV40VLP's may protect cells against apoptosis. Apoptosis was induced inhuman kidney HEK cells by etoposide. As seen in FIG. 3, the addition of50 ng VLPs per 10⁶ cells significantly ameliorated the apoptotic effect,and the cells appeared similar to the untreated control. Similar resultswere obtained with CV-1 cells.

SV40 Targets the Kidney.

The natural environment of SV40 is the primate kidney. It is thereforenot surprising that the virus targets the kidneys in mice. As shown byFIGS. 4A and 4B, extensive staining can be seen with anti-VP1 antibodiesin tubuli of mice 48 hours following a single tail vein injection ofSV40 VLP's. Thereby, emphasizing the applicability and feasibility oftreating ARF disorders using SV40 VLP's or capsid proteins.

Example 3 Establishment of a Mouse Model for Acute Kidney Injury (AKI)

The observed effect of SV40 VLP's and capsid proteins on enhancingchaperone levels and the protective effect on apoptosis in kidney cells,encouraged the inventors to further investigate the possible beneficialeffect of SV40 VLP's and capsid proteins on pathologic disordersspecifically related to kidney. The mercury nephrotoxicity mouse modelwas chosen for the initial studies because of its high reproducibility.For establishing the mouse model, BALB/C female mice, 19-21 grams (6-9weeks), were used. HgCl₂, at the designated concentration, wasadministered IP to mice. Blood was drawn from the tail-vein on thedesignated day and urea was measured using Reflotron urea test (Roche).As shown in FIG. 5, the standard deviation of blood urea levels atvarious HgCl₂ concentrations is low. Based on this experiment a dose of6.5 mg/kg HgCl₂ was selected for the studies of the present invention.The mortality rate at this dose in repeated experiments was consistentlyaround 90%.

Protection of Cultured Mouse Tubular Cells from HgCl₂-Induced Apoptosis

In order to examine the effect of VLP's on nephrotoxicity, mouse tubularcells [Haverty, T. P. et al. J. Cell Biol. 107:1359-1368 (1988)] wereused to represent renal tubular epithelial cells affected in vivo duringacute renal failure. As clearly shown in FIGS. 6A and 6B, treatment ofcells with 15 μM HgCl₂ leads to extensive apoptosis, while infectionwith 50 ng VLPs/10⁶ cells provides effective protection. Similar resultswere obtained for HEK and CV-1 cells (not shown).

Example 4 Treatment of Mice by VLPs Increase Survival Over a Wide DoseRange

As demonstrated above, using the AKI animal-model, most animals injectedwith 6.5 mg/kg HgCl₂ die within 3-4 days. These results were confirmedby FIG. 7A, only 2/20 (10%) of the AKI animals survived. In contrast,survival rate in the animals that received VLPs was much higher. FIG. 7Ashows data of 11 animals that received VLPs in 3 injections at doses60-600 micrograms/mouse. Survival rate of this group was 5/11 (45%).These experiments clearly demonstrate that VLPs have therapeutic effectfor AKI.

To further investigate these encouraging preliminary data, the followingthree experimental groups were examined for analyzing the effect ofVLP's on survival: the first group (Sham) received saline instead ofVLP's and PBS instead of HgCl₂, the second group was treated with HgCl₂and VLP's (AKI+VLPs), and the third group was treated with salineinstead of VLPs and with HgCl₂. VLP's (0.3 mg/kg, in saline) wereadministered to mice via the tail vein in 3 consecutive daily injections(0.1 mg/kg each day, on days −3, −2, −1). Mice that did not receive VLPswere injected in parallel with saline. HgCl₂ in PBS was injected IP onthe fourth day, counted as day 0 in the experiments shown below. Shamanimals received PBS in parallel. The mice were observed for survivalfor 14 additional days. As shown by FIG. 7C, the treatment led to anincrease in survival rate from 12% (6/49) to 63% (19/30). Statisticalanalysis using Kaplan-Meier/Mantel-Cox log-rank test indicatedsignificance at P=0.000002. As shown by FIG. 7B, the attenuating effectwas seen over a wide range of doses, from 3 μg/kg to 3 mg/kg.

Example 5 VP1 Pentamers are Active in Attenuation of AKI

The inventors next examined whether the entire capsids are needed forthe observed beneficial effect, or whether sufficient amount ofinformation for protection from apoptosis may be present in the VP1pentamers. SV40 VLP's consist of 72 pentamers of the major capsidprotein VP1 (360 monomers). The structure of the monomers is a β-barrelcore with extending amino and carboxy-terminal arms. The pentamers areheld together via the 360 carboxy-terminal arms. VP1ΔC, a mutated VP1capsid protein deleted in the carboxy terminal arm, retains the corestructure while it cannot assemble into VLPs. The deletion mutantVP1ΔC65-His [Roitman-Shemer, V. et al. Biochem Biophys. Res. Commun.353:424-430 (2007)] was produced in Sf9 cells and purified on Ni-NTAresin (Qiagen) via the His-tag. Coomassie-stained SDS-PAGE (FIG. 8A)shows, in the lanes loaded with 1 mg protein, that the protein is about95% pure.

Survival experiments were conducted as described above for VLPs, onsmall groups of mice (4 animals in each group). As shown by FIG. 8B, VP1treatment resulted in 75% (¾) survival in mice injected with 0.750mg/kg. These results indicated that VP1ΔC is also effective inprotection against mercury insult, although at a higher dose whencompared to VLPs.

Example 6 Effect of SV40 VLP's and Capsid Proteins on Different DiseaseParameters

The inventors further investigated the applicability and feasibility oftreating pathologic disorders, preferably, ARF, using SV40 VLP's, onadditional kidney failure parameters. One of the parameters examined waskidney structure and morphology. Representative kidneys of the 3 groupsof mice harvested on day 3 following the mercury insult are shown below.While the AKI kidneys were smaller and pale, kidneys from VLP-treatedmice appeared similar to sham kidneys (FIG. 9A). The beneficial effectof VLP's was also observed when additional parameters were examined. Asshown by FIG. 9B, at 4 days post infection, the increase of both serumurea and creatinine levels, two hallmarks of kidney failure, wassignificantly attenuated in VLP-treated AKI mice in comparison tonon-treated AKI mice. Of note, serum urea in the AKI mice increasedcontinuously during the 4 days of the experiment. In contrast, as shownby FIG. 9C, serum urea in the VLP-treated AKI animals reached a peak onday 3 and decreased thereafter. This pattern is compatible with recoveryof the renal function of the VLP-treated animals and correlated with ahigher survival rate.

Example 7 The mechanism of Protection

In an attempt to analyze the possible mechanism of protection of AKIanimals by SV40 VLP's and VP1 capsid protein, the involvement of stresssignals was next examined.

Level of lipid peroxidation was measured by the thiobarbituric acidreactive substances (TBARS) assay [Esterbauer, H. and Cheeseman, K. H.Methods Enzymol. 186:407-421 (1990)]. The level was reduced in theVLP-treated mice (FIG. 10), suggesting that attenuation of oxidativestress by VLPs mediates, at least partially, the protection of renalfunction from mercury-induced nephrotoxicity.

In a parallel study (unpublished data, not shown), the inventors reveledthat treatment of cells using VP1 VLPs, leads to a very rapid activationof PLC-γ and its signaling pathway. In particular, rapid activation (at1 hour) of PARP-1 and caspases 3 and 10, suggesting initiation ofapoptotic pathway. However the cell do not progress to apoptosis, andthe process is stopped by the activation, at 6 hours, of chaperones(Hsp/c70) and the PI3K-PKB/Akt survival pathway, includingphosphorylation of PI3K-PKB/Akt and Bad proteins. Therefore, withoutbeing bound by any theory, the inventors hypothesis that the SV40 VP1capsid protein (or any mutant or VLP thereof) induces the PI3K-PKB/Aktsurvival pathway.

The invention claimed is:
 1. A pharmaceutical composition, in dosageform, comprising a pharmaceutically acceptable carrier and an activeingredient, wherein said active ingredient consists of at least onecapsid protein selected from the group consisting of (1) a wild-typeSV40 VP1 capsid protein or pentamer thereof, (2) a 65 amino acidtruncated mutant of the SV40 VP1 capsid protein deleted in its carboxyterminal arm, and (3) a virus-like particle (VLP) consisting of at leastone wild-type SV40 VP1 capsid protein of (1) or truncated mutant of (2),wherein the composition is DNA-free.
 2. The composition according toclaim 1, wherein said capsid protein comprises at least one wild-typeSV40 VP1 capsid protein or pentamer thereof or a VLP consisting of saidwild-type SV40 VP1 capsid protein.
 3. The composition in accordance withclaim 2, wherein said capsid protein consists of a virus-like particle(VLP) consisting of wild-type SV40 VP1 capsid protein.
 4. Thecomposition according to claim 2, wherein said capsid protein consistsof a pentamer consisting of wild-type SV40 VP1 capsid protein.
 5. Thecomposition according to claim 1, wherein said capsid protein consistsof a wild-type SV40 VP1 capsid protein.
 6. A pharmaceutical compositionfor rescuing cells from any one of oxidative stress and apoptosis andfor inducing survival pathways in said cells, wherein said cells are ofa subject suffering from ARF (Acute Renal Failure), said compositionbeing in dosage form and comprising a pharmaceutically acceptablecarrier and an active ingredient, wherein said active ingredientconsists of a therapeutically-effective amount of at least one capsidprotein selected from the group consisting of (1) a wild-type SV40 VP1capsid protein or pentamer thereof, (2) a 65 amino acid truncated mutantof a SV40 VP1 capsid protein deleted in its carboxy terminal arm, and(3) a virus-like particle (VLP) consisting of at least one wild-typeSV40 VP1 capsid protein of (1) or said truncated mutant of (2), whereinsaid composition is DNA-free.
 7. The composition according to claim 6,wherein said capsid protein is a wild-type SV40 VP1 capsid protein orpentamer thereof, or a virus-like particle (VLP) consisting of at leastone wild-type SV40 VP1 capsid protein.
 8. A pharmaceutical composition,in dosage form, comprising a pharmaceutically acceptable carrier and atleast one capsid protein consisting of a 65 amino acid truncated mutantof a wild-type SV40 VP1 capsid protein deleted in its carboxy terminalarm or a virus-like particle (VLP) consisting of at least one saidtruncated mutant.
 9. A pharmaceutical composition for rescuing cellsfrom any one of oxidative stress and apoptosis and for inducing survivalpathways in said cells, wherein said cells are of a subject sufferingfrom ARF (Acute Renal Failure), said composition comprising apharmaceutically acceptable carrier and, as an active ingredient, atherapeutically-effective amount of at least one 65 amino acid truncatedmutant of a wild-type SV40 VP1 capsid protein deleted in its carboxyterminal arm or a virus-like particle (VLP) consisting of at least onesaid truncated mutant.
 10. A method for treatment of Acute Renal Failure(ARF) comprising administering to a subject in need thereof, as anactive ingredient, a therapeutically-effective amount of at least onewild-type SV40 VP1 capsid protein, or a 65 amino acid truncated mutantof the SV40 VP1 capsid protein deleted in its carboxy terminal arm, orvirus-like particles (VLPs) comprising at least one wild-type SV40 VP1capsid protein or the truncated mutant, or a composition comprising thesame.
 11. The method according to claim 10, wherein said activeingredient is at least one wild-type SV40 VP1 capsid protein or at leastone virus-like particle (VLP) or pentamer comprising the wild-type SV40VP1 capsid protein.
 12. The method according to claim 11, wherein saidARF is AKI (Acute Kidney Injury).
 13. The method according to claim 10,wherein said ARF is AKI (Acute Kidney Injury).
 14. A method fortreatment of ARF (Acute Renal Failure) comprising administering to asubject in need thereof a therapeutically-effective amount of at leastone wild-type SV40 VP1 capsid protein or a composition comprising thesame.
 15. The method according to claim 14, wherein said ARF (AcuteRenal Failure) is AKI (Acute Kidney Injury).
 16. A method for treatmentof ARF (Acute Renal Failure) comprising a step of administering to asubject in need thereof a therapeutically-effective amount of at leastone virus-like particle (VLP) or pentamer comprising a wild-type SV40VP1 capsid protein or a composition comprising the same.
 17. The methodaccording to claim 16, wherein said ARF (Acute Renal Failure) is AKI(Acute Kidney Injury).
 18. A method for rescuing cells from oxidativestress or apoptosis, and inducing survival pathways in said cells,wherein said cells are cells of a subject suffering from ARF (AcuteRenal Failure), comprising contacting cells undergoing oxidative stressor apoptotic process, with an effective amount of at least one wild-typeSV40 VP1 capsid protein, or a 65 amino acid truncated mutant of the SV40VP1 capsid protein deleted in its carboxy terminal arm, or virus-likeparticles (VLPs) comprising at least one wild type SV40 VP1 capsidprotein or the truncated mutant, or a composition comprising the same.19. The method according to claim 18, wherein the ARF is AKT (AcuteKidney Injury).